Functional studies on the candidate ATPase domains of Saccharomyces cerevisiae MutLα

P. T. Tran, Robert (Mike) Liskay

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Saccharomyces cerevisiae MutL homologues Mlh1p and Pms1p form a heterodimer, termed MutLα, that is required for DNA mismatch repair after mismatch binding by MutS homologues. Recent sequence and structural studies have placed the NH2 termini of MutL homologues in a new family of ATPases. To address the functional significance of this putative ATPase activity in MutLα, we mutated conserved motifs tot ATP hydrolysis and ATP binding in both Mlh1p and Pms1p and found that these changes disrupted DNA mismatch repair in vivo. Limited proteolysis with purified recombinant MutLα demonstrated that the NH2 terminus of MutLα undergoes conformational changes in the presence of ATP and nonhydrolyzable ATP analogs. Furthermore, two-hybrid analysis suggested that these ATP-binding-induced conformational changes promote an interaction between the NH2 termini of Mlh1p and Pms1p. Surprisingly, analysis of specific mutants suggested differential requirements for the ATPase motifs of Mlh1p and Pms1p during DNA mismatch repair. Taken together, these results suggest that MutLα undergoes ATP-dependent conformational changes that may serve to coordinate downstream events during yeast DNA mismatch repair.

Original languageEnglish (US)
Pages (from-to)6390-6398
Number of pages9
JournalMolecular and Cellular Biology
Volume20
Issue number17
DOIs
StatePublished - 2000

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Saccharomyces cerevisiae
Adenosine Triphosphatases
DNA Mismatch Repair
Adenosine Triphosphate
Proteolysis
Hydrolysis
Yeasts
MutL Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Functional studies on the candidate ATPase domains of Saccharomyces cerevisiae MutLα. / Tran, P. T.; Liskay, Robert (Mike).

In: Molecular and Cellular Biology, Vol. 20, No. 17, 2000, p. 6390-6398.

Research output: Contribution to journalArticle

Tran, P. T. ; Liskay, Robert (Mike). / Functional studies on the candidate ATPase domains of Saccharomyces cerevisiae MutLα. In: Molecular and Cellular Biology. 2000 ; Vol. 20, No. 17. pp. 6390-6398.
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